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OPA ammonia assay<\/strong> SOLUTIONS: OPA solution: Dissolve 2 g of standard grade o-phtaldialdehyde (P- 1378 Sigma) in 50 ml of ethanol in the dark.<\/p>\n Ammonium chloride stock solution (10mM): Dissolve 535 mg dried NH4Cl in 1000 ml of MilliQ water.<\/p>\n WORKING REAGENT: BCO-DMO Processing Notes<\/strong> BCO-DMO Edits<\/strong>
\nPrepare all solutions in acid-washed (0.1% HCl), triple-rinsed (MilliQ H2O) brown HDPE bottles. Prepare stock solutions with freshly drawn ultra-pure deionized Milli-Q water. For the OPA solution use ethanol of the highest purity available.<\/p>\n
\nSodium sulfite solution: Dissolve 1 g sodium sulfite in 125 ml MilliQ water.
\n\u00a0
\nBorate buffer solution: Dissolve 40 g disodium tetraborate decahydrate in 1000 ml MilliQ water. If necessary, filter the solution to remove turbitity. The solution is stable.<\/p>\n
\nIn a HDPE bottle mix 1000 ml of borate buffer with 50 ml of OPA solution and 5 ml of sodium sulfite. This reagent is light-sensitive. Let the reagent age at least one day before use. Store dark for at least 2 month.
\n\u00a0
\nPROTOCOL:
\nPrepare 2 \u00b5M ammonium standard in a 250ml HDPE bottle: Add 40 \u00b5l of ammonium chloride stock solution (10 mM) to 200 ml Milli-Q water.
\n\u00a0
\nPrepare standard curve as follows:
\nStandard [nM] MilliQ water 2 \u00b5M NH4Cl
\n0 80 0
\n0 80 0
\n10 79.6 0.4
\n25 79 1
\n500 60 20
\n1000 40 40
\n2000 0 80
\n\u00a0
\nFill 80 ml of water sample into a 250-ml HDPE bottle.
\nAdd 20 ml of working reagent to all samples and standards.
\nIncubate for accurately 30 min at 65\u00b0C and cool to room temperature.
\nAnalyze fluorescence intensity in Turner fluorometer with CDOM\/Ammonia kit.
\n\u00a0
\nReference:
\nHolmes, R. M., A. Aminot, R. Kerouel, B. A. Hooker and B. J. Peterson (1999). A simple and precise method for measuring ammonium and marine and freshwater ecosystems. Canadian Journal of Fisheries and Aquatic Sciences 56(10): 1801-1809. Keroul & Aminot (1997) Fluorometric determination of ammonia in sea and estuarine waters by direct segmented flow analysis. Marine Chemistry 57: 265-275.
\n\u00a0
\n\u00a0
Nitrite Determination <\/strong>
\nREAGENTS:
\nSulfanilamide reagent: Add 100 ml conc. HCl to 700ml MilliQ water
\nAdd 10g Sulfanilamide, stir to dissolve.
\nMake up volume to 1000 ml
\nStore at 4?C in the dark, stable for several weeks.
\n\u00a0
\nNaphthylethylenediamine (NED) reagent: Dissolve 1 g of N-(1-Naphthyl)-ethylendiamin-dihydrochloride in 1000 ml of MilliQ water. Store at 4?C in the dark, stable for several weeks.
\n\u00a0
\nNitrite standard stock solution (10 mM): Dissolve 0.690 g dried NaNO2 in 1000 ml MilliQ water.
\n\u00a0
\nPROCEDURE:
\nPrepare 2 \u00b5M nitrite standard in a 250ml HDPE bottle: Add 40 \u00b5l of nitrite standard stock solution (10 mM) to 200 ml artificial seawater (SCM salts).
\n\u00a0
\nPrepare nitrite standard curve as follows:
\nStandard [nM] SCM salts [ml] 2 \u00b5M Nitrite Standard [ml]
\n0 100 0
\n0 100 0
\n10 99.5 0.5
\n25 98.75 1.25
\n500 75 25
\n1000 50 50
\n2000 0 100
\n\u00a0
\nFill 100 ml of sample water into 250ml HDPE bottles.
\nAdd 2 ml of sulfanilamide reagent, swirl and let stand for 1 min.
\nAdd 2 ml of NED reagent and swirl again.
\nIncubate for 10 to 15 min.
\nRead absorbance at 540nm in a 5cm cuvette
\n\u00a0
\nGrasshoff, Kremling, Erhard (1999) Methods of Seawater Analysis, 3 edn. Wiley-VCH
\n\u00a0
Nitrate Assay for Seawater<\/strong>
\nDeionized water and standards
\nFor preparing standard and reagent solutions use deionized water purified by a distilling unit followed by the Millipore Synergy 185 Water System that produces water with 18 M\u03a9 resistance.
\n\u00a0
\nREAGENTS:
\n2% (w\/v) resorcinol solution: prepare fresh daily by dissolving 2.0 g of ACS reagent grade resorcinol crystals in 100 ml Milli-Q water.
\n\u00a0
\nConcentrated sulfuric acid
\nNitrate stock standard solution (10 mM): Dissolve 0.85 g NaNO3 in 1 L of MilliQ water. Store stock solutions in a 1L polyethylene bottle at 4 \u00b0C in a refrigerator.
\nPrepare working standard solutions from serial dilutions of stock solution with artificial sea water (SCM media \u2013 26 g NaCl, 5 g MgSO4\u20227H2O, 5 g MgCl2\u20226H2O, 1.5 g CaCl2\u20222H2O, 0.1 g KBr in 1 L Milli-Q water).
\n\u00a0
\nPROCEDURE:
\nPrepare 40 \u00b5M nitrate standard in a 250ml HDPE bottle by adding 400 \u00b5l of nitrate standard stock solution (10 mM) to 99.6 ml of artificial seawater (SCM media).
\n\u00a0
\nPrepare nitrate standard curve as follows:
\nStandard [\u00b5M] SCM salts [ml] 40 \u00b5M Nitrate Standard [ml]
\n0 20 0
\n0 20 0
\n0.25 19.875 0.125
\n0.5 19.75 0.25
\n10 15 5
\n20 10 10
\n40 0 20
\n\u00a0
\nTransfer 20.0 ml of a seawater sample or standard into a 250 ml brown PE bottle.
\nAdd 2.4 ml of 2% resorcinol solution and swirl to mix the resorcinol with the sample.
\nCarefully add 20.0 ml of concentrated sulfuric acid, close the bottle with its lid and then gently swirl to mix the solution. Let the bottles stand for 30 min.
\nPlace the bottles in a water bath until it reaches room temperature.
\nMeasure the absorbance of the sample at 505 nm in a 5 cm cuvette against a blank with acidified seawater.
\n\u00a0
\nReference:
\nZhang, J.Z. & Fischer, C.J., 2006. A simplified resorcinol method for direct spectrophotometric determination of nitrate in seawater. Mar. Chem. 99, 220-226.<\/p>\n
\nGenerated from original .xls file \"Ingalls_HC_Data for NODC.xls\" contributed by Anitra Ingalls<\/p>\n
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\n- Cruise Id and station metadata added to each data record
\n- Parameters edited to conform to BCO-DMO parameter naming convention
\n- \"Date (euro)\" removed
\n- Decimal data values padded to consistent decimal places as reported
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